The Developmental Origins of Health and Disease (DOHaD) hypothesis argues that early life events (even pre-conception events) influence the risk of adult disease. Pre- and post-natal environmental factors play vital roles in adult disease risk, ranging from metabolic and cardiovascular disease risk factors to cognitive and behavioral abnormalities and some forms of cancer. Considerable evidence indicates that maternal physical activity dramatically impacts fetal and offspring physiology. The offspring of exercised rodent dams or human mothers have altered body composition, organ weights, metabolism and/or behavioral patterns. Importantly, evidence exists that the maternal/intrauterine environment can have profound effects on multiple generations of offspring. In humans, remarkable findings of grandparent- grandchild associations of disease risk and mortality have raised our awareness of the importance of the intrauterine environment, but little has been done to fully test the extent of the influence on future generations or potential mechanisms. Although it is very well accepted that physical activity is a powerful environmental stimulus, no studies have examined if maternal physical activity during pregnancy affects gene expression or metabolism in offspring over multiple generations. Transgeneration exercise effects have been observed for body composition in rodents, but more in-depth and sophisticated studies have not been performed. In the present application, we propose to test whether "physical activity ancestry" results in changes in body composition, gene expression, metabolism, and DNA methylation in multiple generations of mice offspring. Our extensive Preliminary Data support these Specific Aims: Specific Aims: Aim #1 - Single Generation Offspring Analysis: Determine the impact of maternal exercise before and during pregnancy on body composition, tissue gene expression, metabolism, and DNA methylation in mouse offspring. Offspring in generation F1 will be used for extensive phenotype measurement to determine differences between animals with physical activity ancestry compared to sedentary ancestry. Importantly, all F1 offspring will be sedentary. Aim #2 - Transgeneration Analysis: Determine the impact of maternal exercise before and during pregnancy on body composition, muscle force, tissue gene expression, metabolism, and DNA methylation in multiple generations of mouse offspring. Offspring (16 wk) in generations F1 and F2 will be assessed for body composition, metabolic phenotypes (e.g., glucose tolerance test, free fatty acids, etc.), and gene expression and DNA methylation, with comparisons between animals with physical activity ancestry compared to sedentary ancestry. Importantly, all F2 offspring will be sedentary. PUBLIC HEALTH RELEVANCE: Considerable evidence indicates that maternal physical activity before and during pregnancy dramatically impacts fetal and offspring physiology, with offspring exhibiting altered body composition, organ weights, metabolism and/or behavioral patterns. Transgeneration exercise effects have been observed for body composition in rodents, but more in-depth and sophisticated studies have not been performed. In the present application, we propose to test whether "physical activity ancestry" results in changes in body composition, gene expression, metabolism, and DNA methylation in multiple generations of mouse offspring.